CA2195303A1 - Method for selecting high-expressing host cells - Google Patents

Method for selecting high-expressing host cells

Info

Publication number
CA2195303A1
CA2195303A1 CA002195303A CA2195303A CA2195303A1 CA 2195303 A1 CA2195303 A1 CA 2195303A1 CA 002195303 A CA002195303 A CA 002195303A CA 2195303 A CA2195303 A CA 2195303A CA 2195303 A1 CA2195303 A1 CA 2195303A1
Authority
CA
Canada
Prior art keywords
gene
product
host cells
selectable
donor site
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CA002195303A
Other languages
French (fr)
Other versions
CA2195303C (en
Inventor
Craig W. Crowley
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Genentech Inc
Original Assignee
Craig W. Crowley
Genentech, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Craig W. Crowley, Genentech, Inc. filed Critical Craig W. Crowley
Publication of CA2195303A1 publication Critical patent/CA2195303A1/en
Application granted granted Critical
Publication of CA2195303C publication Critical patent/CA2195303C/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6456—Plasminogen activators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/715—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
    • C07K14/7151—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons for tumor necrosis factor [TNF], for lymphotoxin [LT]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C07K16/4283—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an allotypic or isotypic determinant on Ig
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/67—General methods for enhancing the expression
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2830/00—Vector systems having a special element relevant for transcription
    • C12N2830/42—Vector systems having a special element relevant for transcription being an intron or intervening sequence for splicing and/or stability of RNA
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2840/00—Vectors comprising a special translation-regulating system
    • C12N2840/20—Vectors comprising a special translation-regulating system translation of more than one cistron
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2840/00—Vectors comprising a special translation-regulating system
    • C12N2840/44—Vectors comprising a special translation-regulating system being a specific part of the splice mechanism, e.g. donor, acceptor

Abstract

A method for selecting recombinant host cells expressing high levels of a desired protein is described. This method utilizes eukaryotic host cells harboring a DNA construct comprising a selectable gene (preferably an amplifiable gene) and a product gene provided 3' to the selectable gene. The selectable gene is positioned within an intron defined by a splice donor site and a splice acceptor site and the selectable gene and product gene are under the transcriptional control of a single transcriptional regulatory region. The splice donor site is generally an efficient splice donor site and thereby regulates expression of the product gene using the transcriptional regulatory region.
The transfected cells are cultured so as to express the gene encoding the product in a selective medium comprising an amplifying agent for sufficient time to allow amplification to occur, whereupon either the desired product is recovered or cells having multiple copies of the product gene are identified.
CA002195303A 1994-08-05 1995-07-28 Method for selecting high-expressing host cells Expired - Lifetime CA2195303C (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US08/286,740 US5561053A (en) 1994-08-05 1994-08-05 Method for selecting high-expressing host cells
US08/286740 1994-08-05
PCT/US1995/009576 WO1996004391A1 (en) 1994-08-05 1995-07-28 Method for selecting high-expressing host cells

Publications (2)

Publication Number Publication Date
CA2195303A1 true CA2195303A1 (en) 1996-02-15
CA2195303C CA2195303C (en) 2008-01-15

Family

ID=23099965

Family Applications (1)

Application Number Title Priority Date Filing Date
CA002195303A Expired - Lifetime CA2195303C (en) 1994-08-05 1995-07-28 Method for selecting high-expressing host cells

Country Status (13)

Country Link
US (1) US5561053A (en)
EP (1) EP0770136B1 (en)
JP (2) JPH10503376A (en)
AT (1) ATE353970T1 (en)
AU (1) AU704408B2 (en)
CA (1) CA2195303C (en)
DE (1) DE69535389T2 (en)
DK (1) DK0770136T3 (en)
ES (1) ES2282994T3 (en)
IL (1) IL114820A0 (en)
MX (1) MX9700852A (en)
PT (1) PT770136E (en)
WO (1) WO1996004391A1 (en)

Families Citing this family (54)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US4965199A (en) * 1984-04-20 1990-10-23 Genentech, Inc. Preparation of functional human factor VIII in mammalian cells using methotrexate based selection
US6090382A (en) * 1996-02-09 2000-07-18 Basf Aktiengesellschaft Human antibodies that bind human TNFα
SI9720020B (en) 1996-02-09 2001-12-31 Basf Ag Human antibodies that bind human TNF alpha
US5994511A (en) 1997-07-02 1999-11-30 Genentech, Inc. Anti-IgE antibodies and methods of improving polypeptides
US6172213B1 (en) 1997-07-02 2001-01-09 Genentech, Inc. Anti-IgE antibodies and method of improving polypeptides
US20030092002A1 (en) 1997-09-17 2003-05-15 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
US7169906B2 (en) * 1997-09-17 2007-01-30 Genentech, Inc. PRO211 polypeptides
US6974689B1 (en) * 1997-09-18 2005-12-13 Genentech, Inc. Nucleic acid encoding PRO211 polypeptides
US6897066B1 (en) 1997-09-26 2005-05-24 Athersys, Inc. Compositions and methods for non-targeted activation of endogenous genes
WO1999015650A1 (en) * 1997-09-26 1999-04-01 Athersys, Inc. Expression of endogenous genes by non-homologous recombination of a vector construct with cellular dna
US6740503B1 (en) 1997-09-26 2004-05-25 Athersys, Inc. Compositions and methods for non-targeted activation of endogenous genes
JP2002539800A (en) * 1999-03-24 2002-11-26 ジーン セラピー システムズ インコーポレーテッド Method for producing transcriptionally active DNA fragment
CA2385102A1 (en) * 1999-07-12 2001-01-18 Genentech, Inc. Expression vectors and methods
US20050005310A1 (en) * 1999-07-12 2005-01-06 Genentech, Inc. Expression vectors and methods
GB2355010A (en) * 1999-10-04 2001-04-11 Jan Joris Brosens Use of cell-specific splicing in gene therapy
US20020165175A1 (en) * 2001-04-17 2002-11-07 Xiaowu Liang Fast and enzymeless cloning of nucleic acid fragments
NZ543755A (en) 2001-08-29 2008-04-30 Genentech Inc Bv8 nucleic acids and polypeptides with mitogenic activity
EP2022799A2 (en) 2001-11-16 2009-02-11 Biogen Idec Inc. Polycistronic expression of antibodies
US20030157641A1 (en) * 2001-11-16 2003-08-21 Idec Pharmaceuticals Corporation Polycistronic expression of antibodies
JP2006512061A (en) * 2002-11-14 2006-04-13 ジェネンテック・インコーポレーテッド Intron fusion constructs and methods of use for selecting high expression producing cell lines
DE60312039T2 (en) * 2002-12-20 2007-11-08 Chromagenics B.V. MEANS AND METHODS FOR THE PRODUCTION OF A PROTEIN BY CHROMATIN OPENERS WHICH MAY MAKE CHROMATIN ACCESSIBLE TO TRANSCRIPTION FACTORS
EP1615659B1 (en) 2003-03-12 2014-04-16 Genentech, Inc. Use of bv8 and/or eg-vegf to promote hematopoiesis
JP2007529223A (en) * 2004-03-15 2007-10-25 バイオジェン・アイデック・エムエイ・インコーポレイテッド Methods and constructs for expressing polypeptide multimers in eukaryotic cells using alternative splicing
JP5291341B2 (en) 2004-11-08 2013-09-18 クロマジェニックス ベー ヴェー Selection of host cells that express proteins at high levels
US8039230B2 (en) * 2004-11-08 2011-10-18 Chromagenics B.V. Selection of host cells expressing protein at high levels
US20060195935A1 (en) 2004-11-08 2006-08-31 Chromagenics B.V. Selection of host cells expressing protein at high levels
ES2384391T3 (en) 2004-11-08 2012-07-04 Chromagenics B.V. Selection of host cells that express protein at high levels
US8999667B2 (en) * 2004-11-08 2015-04-07 Chromagenics B.V. Selection of host cells expressing protein at high levels
EP1996705B1 (en) * 2006-03-20 2011-08-31 ChromaGenics B.V. Expression augmenting dna fragments, use thereof, and methods for finding thereof
WO2009008837A1 (en) * 2007-07-06 2009-01-15 Temasek Life Sciences Laboratory Limited Lexa-based chemical-inducible gene expression system for use in transgenic animals
CN101802192A (en) 2007-10-15 2010-08-11 中外制药株式会社 Method for production of antibody
WO2010147462A2 (en) 2009-06-15 2010-12-23 Cellagenics B.V. Novel stringent selectable markers
WO2011159157A1 (en) 2010-06-15 2011-12-22 Cellagenics B.V. Novel intergenic elements for enhancing gene expression
US9650630B2 (en) 2010-09-01 2017-05-16 Cellagenics B.V. Nucleic acid fragments from a ribosomal protein promoter for enhancing gene expression
WO2012149197A2 (en) 2011-04-27 2012-11-01 Abbott Laboratories Methods for controlling the galactosylation profile of recombinantly-expressed proteins
US9067990B2 (en) 2013-03-14 2015-06-30 Abbvie, Inc. Protein purification using displacement chromatography
US9334319B2 (en) 2012-04-20 2016-05-10 Abbvie Inc. Low acidic species compositions
WO2013158273A1 (en) 2012-04-20 2013-10-24 Abbvie Inc. Methods to modulate c-terminal lysine variant distribution
WO2013176754A1 (en) 2012-05-24 2013-11-28 Abbvie Inc. Novel purification of antibodies using hydrophobic interaction chromatography
SI2870247T1 (en) * 2012-07-05 2019-10-30 Hoffmann La Roche Expression and secretion system
US9512214B2 (en) 2012-09-02 2016-12-06 Abbvie, Inc. Methods to control protein heterogeneity
KR20150043523A (en) 2012-09-02 2015-04-22 애브비 인코포레이티드 Methods to control protein heterogeneity
SG11201507230PA (en) 2013-03-12 2015-10-29 Abbvie Inc Human antibodies that bind human tnf-alpha and methods of preparing the same
WO2014159579A1 (en) 2013-03-14 2014-10-02 Abbvie Inc. MUTATED ANTI-TNFα ANTIBODIES AND METHODS OF THEIR USE
WO2014151878A2 (en) 2013-03-14 2014-09-25 Abbvie Inc. Methods for modulating protein glycosylation profiles of recombinant protein therapeutics using monosaccharides and oligosacharides
US9017687B1 (en) 2013-10-18 2015-04-28 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same using displacement chromatography
WO2014162318A2 (en) * 2013-03-30 2014-10-09 Usha Biotech Limited Methods and constructs for expressing biologically active proteins in mammalian cells
US9598667B2 (en) 2013-10-04 2017-03-21 Abbvie Inc. Use of metal ions for modulation of protein glycosylation profiles of recombinant proteins
US8946395B1 (en) 2013-10-18 2015-02-03 Abbvie Inc. Purification of proteins using hydrophobic interaction chromatography
US9181337B2 (en) 2013-10-18 2015-11-10 Abbvie, Inc. Modulated lysine variant species compositions and methods for producing and using the same
US9085618B2 (en) 2013-10-18 2015-07-21 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same
WO2015073884A2 (en) 2013-11-15 2015-05-21 Abbvie, Inc. Glycoengineered binding protein compositions
WO2020020471A1 (en) 2018-07-27 2020-01-30 Umc Utrecht Holding B.V. Ascl2-responsive reporters for labeling of intestinal stem cell activity

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4399216A (en) * 1980-02-25 1983-08-16 The Trustees Of Columbia University Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
US4634665A (en) * 1980-02-25 1987-01-06 The Trustees Of Columbia University In The City Of New York Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
FI86885C (en) * 1984-04-20 1992-10-26 Genentech Inc Method for Preparation of Human Recombinant Factor VIII and Nucleic Acid Sequences and Vectors Used thereto
US4965199A (en) * 1984-04-20 1990-10-23 Genentech, Inc. Preparation of functional human factor VIII in mammalian cells using methotrexate based selection
EP0215548B2 (en) * 1985-06-27 1998-01-07 Zymogenetics, Inc. Expression of protein C
DE3730599A1 (en) * 1986-09-12 1988-07-07 Genentech Inc METHOD FOR THE CONTINUOUS PRODUCTION OF HETEROLOGICAL PROTEINS IN EUKARYONTIC HOST CELLS
US5043270A (en) * 1989-03-31 1991-08-27 The Board Of Trustees Of The Leland Stanford Junior University Intronic overexpression vectors
JP3438889B2 (en) * 1991-03-29 2003-08-18 ジェネンテク，インコーポレイテッド Method for selecting a recombinant host cell that expresses a desired protein at a high level
MX9305183A (en) * 1992-08-27 1994-05-31 Us Agriculture PORTABLE INTRON, VIRAL GENOMIC DNA MOLECULA, LIVE VIRUS AND CONTAINING VACCINE AND METHOD FOR OBTAINING IT.

Also Published As

Publication number Publication date
EP0770136B1 (en) 2007-02-14
PT770136E (en) 2007-05-31
WO1996004391A1 (en) 1996-02-15
CA2195303C (en) 2008-01-15
EP0770136A1 (en) 1997-05-02
ES2282994T3 (en) 2007-10-16
DE69535389D1 (en) 2007-03-29
US5561053A (en) 1996-10-01
IL114820A0 (en) 1995-12-08
JP4528940B2 (en) 2010-08-25
AU704408B2 (en) 1999-04-22
MX9700852A (en) 1997-04-30
JP2006296438A (en) 2006-11-02
DE69535389T2 (en) 2007-10-31
ATE353970T1 (en) 2007-03-15
DK0770136T3 (en) 2007-06-11
AU3204595A (en) 1996-03-04
JPH10503376A (en) 1998-03-31

Similar Documents

Publication Publication Date Title
CA2195303A1 (en) Method for selecting high-expressing host cells
ES8507176A1 (en) Method for producing a recombinant baculovirus expression vector.
WO1995027071A3 (en) An adenovirus supervector system
WO1998045456A8 (en) 2-deoxyglucose-6-phosphate (2-dog-6-p) phosphatase dna sequences as selection markers in plants
AU560340B2 (en) Rennin from recombinant dna
WO1998042851A8 (en) Plants with modified growth
DK0719343T3 (en) Adeno-associated viral (AAV) liposomes and method associated therewith
WO1984002921A3 (en) Glucoamylase cdna
WO1996037623B1 (en) Retroviral vectors
EP0827410A4 (en) Method for introducing and expressing genes in animal cells
MX9709549A (en) Recombinant adenoviruses, use thereof for preparing aavs, complementary cell line, and pharmaceutical compositions containing said adenoviruses.
UA42675C2 (en) Method for gene activation of eucariotic cell line genom, which is transcriptionally latent in normal conditions, method for modification of gene expression characteristics for eucariotic cell line gene, differentiated eucariotic cell line (variants), dna-constract
CA2311643A1 (en) Compositions and methods for inducing gene expression
AU4200897A (en) Novel host cells and methods of producing proteins
IE862178L (en) Poly-kringle plasminogen activator
AU716500B2 (en) Nucleic acid constructs for the cell cycle-regulated expression of genes, cells containing such constructs and their use for preparing drugs
IE840258L (en) Expression of gene products.
WO1992017566A3 (en) Methods for selection of recombinant host cells expressing high levels of a desired protein
IL90334A0 (en) Genetic transformation of b.thuringiensis,vectors used therefor,host cells transformed by the vectors and use of said cells as insecticides
AU5961596A (en) Method for expression and isolation of biologically active m olecules in urine
CA2102594A1 (en) Gene manipulation and expression using genomic elements
EP0319206A3 (en) Gene amplification
EP0154566A3 (en) Viral enhancer dna segments
CA2337088A1 (en) Methods and constructs for protein expression
WO1998039427A3 (en) Gene therapy using bone marrow transplants transfected with therapeutic genes under the control of tissue-specific promoters

Legal Events

Date Code Title Description
EEER Examination request
MKEX Expiry

Effective date: 20150728